中间普氏菌培养上清对牙龈成纤维细胞RANKL／OPG表达的影响

目的：探讨中间普氏菌培养上清对牙龈成纤维细胞核因子-κB受体活化因子配体（receptor activator for nuclear factor-κB ligand，RANKL）、骨保护素（osteoprotegerin，OPG）表达的影响。方法：将系列浓度的中间普氏菌培养上清作用于牙龈成纤维细胞，显微镜下观察细胞的变化。实时定量 RT-PCR法和 Western blot法分别检测 RANKL、OPG mRNA及蛋白水平的表达，并计算 RANKL/OPG的比值。结果：牙龈成纤维细胞的数量随中间普氏菌培养上清浓度的增加而减少，50μg/mL 时，显微镜下没有活细胞存在。在mRNA水平上和蛋白水平上，RANKL/OPG的比值在12．5μg/mL处理组均高于对照组，差异具有统计学意义（P＜0．01）。结论：中间普氏菌培养上清可诱导人牙龈成纤维细胞表达 RANKL 和 OPG，破坏 RANKL/OPG比例的平衡，影响破骨细胞分化的细胞因子微环境，在破骨细胞分化和牙周炎骨吸收中起着一定的调节作用。

Effect of Prevotella intermedia supernatant on the expression of RANKL and OPG in human gingival fibroblasts

Objective：To evaluate the effects of Prevotella intermedia supernatant on the expression of receptor activator of NF-κB ligand （RANKL）and osteoprotegerin （OPG）in human gingival fibroblasts （HGFs）.Methods：The HGFs were treated with gradient Prevotella intermedia supernatant.HGFs were observed under the microscope.The real-time RT-PCR and Western blot analysis were used for testing the expression of RANKL and OPG,and the RANKL/OPG ratio was calculated.Results：The number of cells decreased with increase of the supernatant concentration.At 50 μg/mL,no cell survival could be observed under the microscope.The ratio of RANKL/OPG was increased in the cells treated with 12.5 μg/mL of Prevotella intermedia supernatant（P〈0.01）.Conclusions：Prevotella intermedia supernatant could disrupt the homeostasis of RANKL/OPG,destroy the microenvironment of osteoclast differentiation. Prevotella intermedia supernatant may play a regulatory role in osteoclast differentiation and bone resorption in periodontitis.